ber is a gene which exists in the human twenty-second chromosome and abl is a gene which exists in the human ninth chromosome, and Philadelphia chromosome is formed by translocation of the human twenty-second and ninth chromosomes. It is known that a gene product of the chromosome, BCR-ABL, is a protein having tyrosine kinase activity and constantly generates the growth signal to cause aberrant growth of cells (see, for example, Non-Patent Document 2).
of Cell counting Kit-8 (5 mmol/l WST-8, 0.2 mmol/1 1-Methoxy PMS, 150 mmol/l NaCl) (manufactured by Dojindo) was added to each well. After reaction for color development in a CO2 incubator for 3 hours, an absorbance of formazan, generated by reduction of WST-8 was determined at 450 nm using Multi-level counter ARVOsx (manufactured by Wallac).
In the RPMI-1640/FCS medium containing 0.1% DMSO, when absorbance of a region in which cells after culturing in the CO2 incubator for 72 hours were seeded is defined as a cell growth inhibition rate of 0% and absorbance of a region in which cells were not seeded is defined as a cell growth inhibition rate of 100%, a log conc value in terms of log (inhibition rate/(100−inhibition rate)) and a plotted IC50 value (μM) were calculated. The results are shown in Table 3.
As a control drug, 4-(4-methylpiperazin-1-ylmethyl)-N-{4-methyl-3-[4-(3-pyridyl)pyrimidin-2-ylamino]phenyl}benzamide (see Patent Document 1) was used.
TABLE 3RatioK562 cellsU937 cells(U937 cells/Test drugs(IC50 value: μM)(IC50 value: μM)K562 cells)Example 20.0028155357Example 40.00202814000Example 60.00124.33583Example 80.000984.64694Example 100.00218.13857Example 120.00564.7839Example 140.00792.0253Example 160.00114.13727Example 180.00484.6958
Therefore, inhibition of the BCR-ABL tyrosine kinase activity makes it possible to suppress cell growth caused by the kinase and a compound which inhibits the activity is suited for use as a therapeutic agent for diseases such as chronic myelogenous leukemia, acute lymphoblastic leukemia and acute myelogenous leukemia. Although Glivec® (see, for example, Patent Document 1) has already been put on the market as a drug having the same action, other drugs having the same action mechanism have never been put on the market and thus it has been required to develop more excellent medicines.
It has recently been reported that recurrence is often recognized in patients wherein remission is attained by administration of Glivec® in BCR-ABL-positive acute lymphoblastic leukemia, in addition to examples of blastic crisis of chronic myelogenous leukemia (see, for example, Non-Patent Document 3). As a result of examination of leukemia cells of the patients suffering from the recurrence of disease, the appearance of a variant such as E255K is recognized (see, for example, Non-Patent Documents 4 to 7). Also in examples of administration of Glivec® to the patients with BCR-ABL-positive acute lymphoblastic leukemia, the appearance of resistant cells which mainly exhibits variation of E255K is recognized (see, for example, Non-Patent Document 8). With an increase in use of Glivec®, resistant patients further increase and thus it is required to develop a therapy.    Patent Document 1: Japanese Unexamined Patent No. 6-87834    Patent Document 2: Pamphlet of International Publication WO 02/22597    Non-Patent Document 1: Shtivelman E, et al.: Nature, 1985, 315, 550-554    Non-Patent Document 2: Daley G Q, et al.: Science, 1990, 247, 824-830    Non-Patent Document 3: Druker B J, et al.: N Engl J Med, 2001, 344, 1038-1042    Non-Patent Document 4: Weisberg E, et al.: Drug Resist Updat, 2001, 4, 22-28    Non-Patent Document 5: Gorre M E, et al.: Science, 2001, 293, 876-880    Non-Patent Document 6: Blagosklonny M V: Leukemia, 2002, 16, 570-572    Non-Patent Document 7: Hochhaus A, et al.: Leukemia, 2002, 16, 2190-2196    Non-Patent Document 8: Hofmann W-K, et al.: blood, 2002, 99, 1860-1862    Non-Patent Document 9: Deninger W N, et al.: blood, 2000, 96, 3343-3356    Non-Patent Document 10: J. Org. Chem., 1996, 61, 1133-1135    Non-Patent Document 11: J. Org. Chem., 2000, 65, 1144-1157    Non-Patent Document 12: Recl. Trav. Chim. Pays-Bas., 1950, 69, 673-699    Non-Patent Document 13: J. Heterocycl. Chem., 1970, 7, 1137-1141    Non-Patent Document 14: J. Am. Chem. Soc., 1999, 121, 4369-4378    Non-Patent Document 15: Tetrahedron Lett., 1997, 38, 8005-8008    Non-Patent Document 16: J. Med. Chem., 2002, 45, 3406-3417    Non-Patent Document 17: J. Med. Chem., 2000, 43, 3895-3905    Non-Patent Document 18: J. Med. Chem., 2000, 43, 1508-1518    Non-Patent Document 19: J. Med. Chem., 1975, 18, 1077-1088    Non-Patent Document 20: Bioorg. Med. Chem. Lett., 2001, 11, 2235-2239    Non-Patent Document 21: J. Heterocyclic Chem., 2000, 37, 1457-1462